Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy

• Verfasst von: Keller, Philipp J. [VerfasserIn]
• Verfasst von: Schmidt, Annette D. [VerfasserIn]
• Verfasst von: Wittbrodt, Joachim [VerfasserIn]
• Verfasst von: Stelzer, Ernst H. K. [VerfasserIn]
• Titel: Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy
• Verf.angabe: Philipp J. Keller, Annette D. Schmidt, Anthony Santella, Khaled Khairy, Zhirong Bao, Joachim Wittbrodt & Ernst H. K. Stelzer
• Umfang: 6 S.
• Fussnoten: Gesehen am 02.06.2017
• Titel Quelle: Enthalten in: Nature methods
• Jahr Quelle: 2010
• Band/Heft Quelle: 7(2010), 8, S. 637-642
• ISSN Quelle: 1548-7105
• DOI: doi:10.1038/nmeth.1476
• Datenträger: Online-Ressource
• Sprache: eng
• K10plus-PPN: 155943242X

Abstract

Recording light-microscopy images of large, nontransparent specimens, such as developing multicellular organisms, is complicated by decreased contrast resulting from light scattering. Early zebrafish development can be captured by standard light-sheet microscopy, but new imaging strategies are required to obtain high-quality data of late development or of less transparent organisms. We combined digital scanned laser light-sheet fluorescence microscopy with incoherent structured-illumination microscopy (DSLM-SI) and created structured-illumination patterns with continuously adjustable frequencies. Our method discriminates the specimen-related scattered background from signal fluorescence, thereby removing out-of-focus light and optimizing the contrast of in-focus structures. DSLM-SI provides rapid control of the illumination pattern, exceptional imaging quality and high imaging speeds. We performed long-term imaging of zebrafish development for 58 h and fast multiple-view imaging of early Drosophila melanogaster development. We reconstructed cell positions over time from the Drosophila DSLM-SI data and created a fly digital embryo.